Colorectal cancer (CRC) progression is strongly influenced by interactions between tumor cells and the collagen-rich extracellular matrix. Discoidin domain receptor 1 (DDR1), a collagen-activated receptor tyrosine kinase, has been associated with poor prognosis in several epithelial malignancies, including CRC. However, the mechanisms by which DDR1 contributes to metastatic initiation remain incompletely understood.Here, we investigated the role of DDR1 in regulating CRC cell migration and invasion within type I collagen–rich microenvironments. DDR1 overexpression in low-invasive HT-29 cells was sufficient to enhance collective spheroid migration on collagen. Conversely, pharmacological inhibition or genetic silencing of DDR1 in highly invasive HCT-116 cells significantly impaired two-dimensional and three-dimensional migration as well as invasion within fibrillar collagen matrices.Mechanistically, DDR1 inactivation restored epithelial features, as evidenced by increased membrane localization of E-cadherin, decreased N-cadherin expression and reduced MT1-MMP expression. These findings indicate that DDR1 promotes invasion through coordinated regulation of epithelial adhesion and localized collagen remodeling rather than global extracellular matrix degradation. Phosphoproteomic profiling further suggested that DDR1 activates a non-canonical pro-migratory signaling network independent of AKT and Wnt/β-catenin pathways, involving modulation of GSK-3, WNK1, and RSK signaling.Collectively, our results identify DDR1 as a critical regulator of collagen-dependent migration and invasion in CRC and highlight its kinase activity as a potential therapeutic target to limit metastatic progression.

Objectif L’intolérance aux protéines lysinuriques (LPI) est une maladie autosomique récessive due à un défaut du transport transmembranaire des acides aminés dibasiques. Cette anomalie entraîne un trouble secondaire du cycle de l’urée, exposant à des complications sévères, notamment pulmonaires, hématologiques, immunologiques et rénales, avec des atteintes tubulaires et, plus rarement, glomérulaires pouvant évoluer vers l’insuffisance rénale terminale. Des travaux récents suggèrent que l’hyperéchogénicité colique fœtale pourrait constituer un marqueur échographique précoce du diagnostic de LPI. Méthodes Les patients ont été sélectionnés dans sept centres français de référence des maladies métaboliques héréditaires sur une période de 20 ans (2001–2021). Ont été inclus les patients diagnostiqués avec une LPI et ayant bénéficié d’une échographie prénatale au troisième trimestre, avec recherche de la présence ou non d’une hyperéchogénicité colique fœtale. Un questionnaire standardisé a permis de recueillir rétrospectivement les données cliniques et d’imagerie, incluant antécédents familiaux, âge au diagnostic, données génétiques et biologiques confirmant la LPI, ainsi que le suivi clinique (atteinte rénale, prise en charge thérapeutique). Résultats Entre 2001 et 2021, 38 cas de LPI ont été recensés. L’âge médian au diagnostic était de 5 ans et 9 mois, avec un sex-ratio H/F de 2,1. La consanguinité était présente dans 74,3 % des cas (26/35). Parmi les 15 cas disposant de données échographiques, une hyperéchogénicité colique fœtale a été observée dans 53,3 % (8/15). Conclusion La détection d’une hyperéchogénicité colique fœtale doit alerter les cliniciens, notamment les néphrologues pédiatres, sur la possibilité d’une LPI, et non uniquement d’une cystinurie. La reconnaissance précoce de ce signe échographique prénatal pourrait permettre une prise en charge ciblée dès la naissance, évitant ainsi des complications métaboliques sévères telles que l’hyperammoniémie.

OBJECTIVES:Reliable biomarkers may allow early risk stratification for mortality in older adults. Post-translational modification-derived products (PTMDPs), such as homocitrulline, and the tissue accumulation of advanced glycation end products (AGEs), measured by skin autofluorescence, have been suggested as potential biomarkers of adverse outcomes in older and frail individuals. The aim of this study was to assess their association with 3-year mortality following hospitalisation. METHODS:We followed a cohort of 250 hospitalised patients over 3 years. Serum PTMDPs (homocitrulline, carboxymethyllysine, pentosidine, and MG-H1) were measured using liquid chromatography coupled with tandem mass spectrometry, and AGEs were quantified by skin autofluorescence. We examined associations with mortality using bivariable analyses and a multivariable Cox regression model. An exploratory cut-off for homocitrulline was established at the third quartile of its distribution within the study population. RESULTS:During follow-up, 92 deaths occurred. In bivariable analyses, all biomarkers were associated with mortality. In the multivariable model, homocitrulline levels of >400 μmol/mol Lys were significantly associated with mortality (hazard ratio 1.73, 95 % confidence interval 1.06-2.82), after adjustment for age, frailty, and comorbidities. The model's predictive performance, with an area under the curve (AUC) of 0.76, was only slightly affected by the exclusion of homocitrulline (AUC=0.755). CONCLUSIONS:These results indicate that although elevated homocitrulline was independently associated with mortality, its incremental value for risk prediction was modest. Larger studies are needed to further evaluate the clinical utility of PTMDPs, especially homocitrulline, for risk prediction and clinical decision-making.

Carbamylation is a nonenzymatic post-translational modification that alters protein structural and functional properties and is involved in the pathogenesis of many diseases. It results from isocyanic acid binding to protein amino groups, generating carbamylation-derived products, including homocitrulline (HCit) when the reaction targets the ε-amino group of lysine residues. Isocyanic acid is produced by two major sources in vivo, the spontaneous dissociation of urea and the myeloperoxidase (MPO)-catalyzed conversion of thiocyanate, but their respective contribution to carbamylation is disputed in literature. Here, we compared tissue accumulation of HCit in wild-type versus MPO-deficient mice during ageing. Our results showed that the kinetics and amplitude of carbamylation were not reduced in MPO-deficient mice. Furthermore, carbamylation was intriguingly enhanced in younger MPO-deficient mice, suggesting the presence of compensatory mechanisms. These findings suggest that the MPO pathway is not necessarily required for age-associated systemic carbamylation.

Kidney function was independently associated with free drug concentration; the latter concentration was higher in patients with a lower eGFR. The conformational changes in albumin induced by carbamylation may alter its binding capacity to vitamin K antagonists. The results on the accumulation of free fraction of drugs in patients with low eGFR may be applicable to other drugs that are bound to plasma proteins. The metabolic disturbances associated with CKD might alter drug distribution, decrease albumin drug binding, and thus increase the free (unbound) drug concentrations. Uremic toxins (UTs) can affect the pharmacokinetic or pharmacodynamic activity of certain drugs. Vitamin K antagonists (VKAs) are interesting candidates for the evaluation of potential interactions between UTs and drugs. The primary objective of this study was to investigate the association between free VKA concentrations and the eGFR. Furthermore, we sought to determine whether this relationship was mediated by protein carbamylation (measured by homocitrulline levels) and/or the accumulation of protein-bound UTs (PBUTs). In this prospective cross-sectional study, 389 adult patients treated with VKA were included between May 2021 and June 2023. Levels of free VKAs, total VKAs, homocitrulline, and PBUTs were assayed using liquid chromatography-tandem mass spectrometry. We used a linear regression model to explore the association between kidney function and free VKA levels and mediation analyses to determine whether the association between kidney function and free VKA levels was mediated (at least partly) by PBUTs and/or protein carbamylation. Patients with an eGFR <40 ml/min per 1.73 m 2 or those on chronic hemodialysis had lower total VKA levels, higher free VKA levels, and thus a higher of free/total VKA ratio than those with an eGFR ≥40 ml/min per 1.73 m 2 . Kidney function was independently associated with free VKA levels ( β 1 =0.31 [0.19 to 0.42], P < 0.001). Twenty-one percent (95% confidence interval, 1% to 35%) of the association between kidney function and free VKA levels was mediated by homocitrulline, but not by PBUTs. Our results showed that a low kidney function was associated with an elevation in the VKA free drug fraction. This association was independent of blood albumin levels and appeared to be partly mediated by protein carbamylation.

Protein carbamylation contributes to an increase in the cardiovascular risk in certain patient populations (e.g. in patients with chronic kidney disease (CKD) because of elevated urea concentrations). Homocitrulline (HCit) is a biomarker of overall protein carbamylation. In a study of a large cohort of non-dialysis patients with a confirmed diagnosis of CKD and an estimated glomerular filtration rate (eGFR) <60 mL/min/1.73 m², we sought to determine whether the serum HCit concentration was associated with adverse cardiovascular outcomes and all-cause mortality. CKD-REIN is a prospective cohort of CKD patients with an eGFR <60 mL/min/1.73 m². The baseline serum HCit concentration was centrally measured, using liquid chromatography-tandem mass spectrometry. The 2,195 patients included in the analysis were divided into tertiles (Ts) groups as a function of the baseline HCit concentration (T1 <292, T2 = [292–429], and T3 ≥430 µmol/mol lysine). Cox proportional hazards models were used to estimate hazard ratios (HRs) for the first major adverse cardiovascular event (MACE, defined as a composite of nonfatal stroke, nonfatal myocardial infarction, hospitalization for heart failure, and cardiovascular death) and for death before kidney replacement therapy (KRT, defined as the initiation of chronic dialysis or kidney transplantation). The models were adjusted for baseline comorbidities, laboratory data, and medications. Among the 2,195 included patients, the median [interquartile range (IQR)] age was 68, and the mean eGFR was 34.6 mL/min/1.73 m². The median [IQR] serum HCit was 352 [266–481] µmol/mol lysine. The serum HCit concentration was correlated with the eGFR and the urea concentrations (r = −0.57 and r = 0.73, respectively). In an adjusted linear regression model, the HCit concentration was independently and positively associated with age, eGFR decrease, urea, anemia, and baseline prescription of diuretics, and negatively associated with male sex and a greatly elevated urinary albumin-to-creatinine ratio. The adjusted HR [95% confidence interval] for MACEs as a function of the baseline serum HCit concentration was 1.32 [0.96; 1.83] for patients in T2 and 1.59 [1.13; 2.24] for patients in T3, relative to patients in T1 (Fig. 1). The adjusted HR [95% CI] for death before KRT as a function of the baseline serum HCit concentration was 1.42 [0.99; 2.01] for patients in T2 and 2.01 [1.39; 2.91] for patients in T3, relative to patients in T1 (Fig. 1). Our analysis of a large cohort of patients with CKD demonstrated that the serum HCit concentration was associated with a greater likelihood of a MACE and death. To confirm causality, further studies of therapeutic interventions for preventing or reducing carbamylation are now warranted.

AIM:Advanced glycation end-products (AGEs) are known to play a role in the pathophysiology of type 1 diabetes (T1D) complications. The aim of this study was to assess the predictive value of AGEs indirectly evaluated by skin auto-fluorescence (SAF) on the occurrence of cardiovascular events (CVEs) in T1D. METHODS:We measured baseline SAF in T1D patients with at least 10 years history of diabetes and assessed incident CVEs. An optimum threshold of SAF was determined using ROC curve, and its predictive value was assessed by Cox proportional regression. RESULTS:The study included 179 patients, 53 % of whom were women. At baseline, the mean age was 47.7 ± 15.9 years, the mean duration of diabetes was 26.4 ± 12.2 years. Median HbA1c was 7.7 % (7.3-8.7) and median LDL cholesterol was 2.58 mmol/l (2.14-3.07). Median follow-up was 7.4 years (6.85 - 7.7) with 34 CVEs in 24 patients. The median SAF level was 2.7 (2.3-3.1) in patients with incident CVEs and 2.1 (1.8-2.6) in patients without CVEs. The optimum threshold of SAF to differentiate patients with or without incident CVEs was 2.2. The occurrence of CVE was predicted by the optimum SAF threshold in the unadjusted model (HR 6.46), but also after adjustment with different models (HR 3.15-5.05). CONCLUSION:SAF level is higher in people living with T1D who will present CVEs. Furthermore, SAF threshold of 2.2 predicted the occurrence of CVE. If these results are confirmed, SAF could be a useful marker in cardiovascular risk stratification in T1D.

The importance of HbA1c in the therapeutic monitoring of diabetic patients requires the use of robust and efficient assay methods, especially for point-of-care testing. This study evaluates the analytical performances of the Siemens Healthineers Atellica® DCA. After analyzing linearity and precision, the results were compared with those of the DCA Vantage (Siemens) and the Capillarys 3 Tera (Sebia, capillary electrophoresis). The main interferences (triglycerides and bilirubin) and the effect of total hemoglobin concentration were also assessed. The tests showed a good linearity over a range of 4.5 to 14 % HbA1c, and good precision with coefficients of variation ranging from 1.42 to 2.33 %. The correlation with capillary electrophoresis and the Atellica® DCA was excellent (r² = 0.991 and r² = 0.994, respectively). The accuracy of the results, assessed with external quality samples, was also satisfactory. The method is not sensitive to interference of hypertriglyceridemia up to 10 mmol/L or hyperbilirubinemia (up to 432 μmol/L), and provides reliable results even for low concentrations of total hemoglobin (down to 57,5 g/L). The Atellica® DCA also offers improved ergonomics, with a user-friendly touchscreen and can be connected to the laboratory's management software. this analyzer presents analytical performances suitable for use as point-of-care testing analyzer.

BACKGROUND:Protein carbamylation contributes to an increase in the cardiovascular risk in certain patient populations (e.g. in patients with chronic kidney disease (CKD) because of elevated urea concentrations). Homocitrulline (HCit) is a biomarker of overall protein carbamylation. In a study of a large cohort of non-dialysis patients with a confirmed diagnosis of CKD and an estimated glomerular filtration rate (eGFR) <60mL/min/1.73m2, we sought to determine whether the serum HCit concentration was associated with adverse cardiovascular outcomes and all-cause mortality. METHODS:CKD-REIN is a prospective cohort of CKD patients with an eGFR<60mL/min/1.73m2. The baseline serum HCit concentration was centrally measured. The 2,195 patients included in the analysis were divided into tertiles (T) groups according to the baseline HCit concentration (T1<292, T2=[292-429], and T3≥430 µmol/mol lysine). Adjusted Cox proportional hazards models were used to estimate hazard ratios (HRs) for the first major adverse cardiovascular event (MACE) and death before kidney replacement therapy (KRT). RESULTS:Among the 2,195 included patients, the median age was 68, and the mean eGFR was 34.6 mL/min/1.73m2. The median [IQR] serum HCit was 352[266-481] µmol/mol lysine. The HCit concentration was correlated with the eGFR (r=-0.57) and the urea concentrations (r=0.73). In an adjusted linear regression model, the HCit concentration was independently and positively associated with age, eGFR decrease, urea, anemia, baseline prescription of diuretics, and negatively associated with male sex and an elevated urinary albumin-to-creatinine ratio. The adjusted HR[95%CI] for MACEs as a function of the baseline HCit concentration was 1.32[0.96-1.84] for T2 and 1.63[1.16-2.30] for T3, compared to T1. The risk of death before KRT as a function of the baseline serum HCit concentration was 2.09[1.45-3.03] for T3 and 1.48[1.04-2.11] for T2, compared to T1. CONCLUSIONS:Our analysis of a large cohort of patients with CKD demonstrated that the serum HCit concentration was associated with a greater likelihood of a MACE and death. To confirm causality, further studies of therapeutic interventions for preventing or reducing carbamylation are now warranted.

Background:Diabetic patients experience a significant reduction in life expectancy, primarily due to early cardiovascular complications. A key feature is the premature degradation of elastic fibers (EFs), contributing to vascular stiffness. Objective:This study evaluates the capacity of two antihypertensive agents, minoxidil (a KATP channel opener) and nebivolol (a β-blocker with KATP activity), to restore EF homeostasis and arterial elasticity in diabetic mice. Methods:Mice are treated with two antihypertensive agents: minoxidil (an ATP-sensitive potassium (KATP) channel opener) or nebivolol (a β-blocker also active on KATP channels). The degree of wear and functionality of EF are assessed after these treatments. We complement this analysis by identifying molecular actors from smooth muscle cell cultures. Results:Our data show that by applying these antihypertensive agents in cultured vascular smooth muscle cells in vitro and in diabetic mice, we efficiently stimulate elastogenesis and inhibit elastolysis. Therefore, treatments restore functional EFs and limit their degradation. This brings blood pressure values of diseased mice close to normal ones (as in unaffected mice). Elastogenesis pathway stimulation and elastolysis inhibition are induced by the opening of sensitive KATP channels and the regulation of the forkhead box transcription factor (FOXO1). Conclusion:Minoxidil and nebivolol restore EF integrity and limit vascular aging in diabetic mice via K+ channel opening and FOXO1 repression. These findings highlight potassium channel-FOXO1 signaling as a therapeutic axis to counteract diabetic vascular complications.

The importance of HbA1c in the therapeutic monitoring of diabetic patients requires the use of robust and efficient assay methods, especially for point-of-care testing. This study evaluates the analytical performances of the Siemens Healthineers Atellica® DCA. After analyzing linearity and precision, the results were compared with those of the DCA Vantage (Siemens) and the Capillarys 3 Tera (Sebia, capillary electrophoresis). The main interferences (triglycerides and bilirubin) and the effect of total hemoglobin concentration were also assessed. The tests showed a good linearity over a range of 4.5 to 14 % HbA1c, and good precision with coefficients of variation ranging from 1.42 to 2.33 %. The correlation with capillary electrophoresis and the Atellica® DCA was excellent (r² = 0.991 and r² = 0.994, respectively). The accuracy of the results, assessed with external quality samples, was also satisfactory. The method is not sensitive to interference of hypertriglyceridemia up to 10 mmol/L or hyperbilirubinemia (up to 432 μmol/L), and provides reliable results even for low concentrations of total hemoglobin (down to 57,5 g/L). The Atellica® DCA also offers improved ergonomics, with a user-friendly touchscreen and can be connected to the laboratory's management software. this analyzer presents analytical performances suitable for use as point-of-care testing analyzer.

AIM:Vascular smooth muscle cells (VSMCs) are characterized by a considerable plasticity. Their phenotypic switch (from contractile to synthetic) plays a crucial role in the atherosclerotic process, explaining that numerous studies focus on this phenotypic transition. Thus, it is essential to use VSMCs that have been finely phenotyped for experimental purposes. The use of MOVAS cell line is suitable because, unlike primary cells, it is believed that these cells retain their phenotype and avoid cell senescence in culture. This study aimed to assess the phenotype of MOVAS cells over culture passages to ensure that they retained a contractile phenotype, before using them for further investigations. METHODS:The phenotype of MOVAS cells at different culture passages (P3, P5 and P8) was analysed morphologically and by studying the expression of genes that indicate a contractile (Acta2, Myocd and Cnn1) and synthetic (Klf4 and Lgals3) VSMC phenotype by RT-qPCR. Cell stiffness was analysed by atomic force microscopy and cell adhesion and migration. RESULTS:Our results showed that MOVAS cells rapidly changed morphologically and that the gene expression of contractile markers was significantly reduced in favor of markers specific to the synthetic phenotype. These changes were associated with a reduction in cell stiffness and a significant increase in adhesion and migration properties. CONCLUSION:MOVAS cells undergo a transition from contractile to synthetic phenotype with increasing number of passages in vitro, which means that these cells should be used with caution, at a low number of passages, while being regularly characterized.

BACKGROUND:The metabolic disturbances associated with chronic kidney disease might alter drug distribution, decrease albumin drug binding, and thus increase the free (unbound) drug concentrations. Uremic toxins can affect the pharmacokinetic or pharmacodynamic activity of certain drugs. Vitamin K antagonists (VKA) are interesting candidates for the evaluation of potential interactions between uremic toxins and drugs. The primary objective of the present study was to investigate the association between free VKA concentrations and the estimated glomerular filtration rate (eGFR). Furthermore, we sought to determine whether this relationship was mediated by protein carbamylation (measured by homocitrulline levels) and/or the accumulation of protein-bound uremic toxins (PBUTs). METHODS:In this prospective cross-sectional study, 389 adult patients treated with VKA were included between May 2021 and June 2023. Levels of free VKAs, total VKAs, homocitrulline and PBUTs were assayed using liquid chromatography-tandem mass spectrometry. We used a linear regression model to explore the association between kidney function and free VKA levels and mediation analyses to determine whether the association between kidney function and free VKA levels was mediated (at least partly) by PBUTs and/or protein carbamylation. RESULTS:Patients with an eGFR < 40 mL/min/1.73 m2 or those on chronic hemodialysis had lower total VKA levels, higher free VKA levels and thus a higher of free/total VKA ratio than those with an eGFR ≥ 40 mL/min/1.73 m2. Kidney function was independently associated with free VKA levels (β1=0.31 [0.19; 0.42], p<0.001). Twenty one percent [95%CI, 1%-35%] of the association between kidney function and free VKA levels was mediated by homocitrulline, but not by PBUTs. CONCLUSIONS:Our results showed that a low kidney function was associated with an elevation in the free drug fraction. This association was independent of blood albumin levels and appeared to be partly mediated by protein carbamylation.

Dietary advanced glycation end-products (dAGEs) accumulate in organs and are thought to initiate chronic low-grade inflammation (CLGI), induce glycoxidative stress, drive immunosenescence, and influence gut microbiota. Part of the toxicological interest in glycation products such as dietary carboxymethyl-lysine (dCML) relies on their interaction with receptor for advanced glycation end-products (RAGE). It remains uncertain whether early or lifelong exposure to dAGEs contributes physiological changes and whether such effects are reversible or permanent. Our objective was to examine the physiological changes in Wild-Type (WT) and RAGE KO mice that were fed either a standard diet (STD - 20.8 ± 5.1 µg dCML/g) or a diet enriched with dCML (255.2 ± 44.5 µg dCML/g) from the perinatal period for up to 70 weeks. Additionally, an early age (6 weeks) diet switch (dCML→STD) was explored to determine whether potential harmful effects of dCML could be reversed. Previous dCML accumulation patterns described by our group were confirmed here, with significant RAGE-independent accumulation of dCML in kidneys, ileum and colon over the 70-week dietary intervention (respectively 3-fold, 17-fold and 20-fold increases compared with controls). Diet switching returned tissue dCML concentrations to their baseline levels. The dCML-enriched diet had no significative effect on endogenous glycation, inflammation, oxidative stress or senescence parameters. The relative expression of TNFα, VCAM1, IL6, and P16 genes were all upregulated (∼2-fold) in an age-dependent manner, most notably in the kidneys of WT animals. RAGE knockout seemed protective in this regard, diminishing age-related renal expression of TNFα. Significant increases in TNFα expression were detectable in the intestinal tract of the Switch group (∼2-fold), suggesting a higher sensitivity to inflammation perhaps related to the timing of the diet change. Minor fluctuations were observed at family level within the caecal microbiota, including Eggerthellaceae, Anaerovoracaceae and Marinifilaceae communities, indicating slight changes in composition. Despite chronic dCML consumption resulting in higher free CML levels in tissues, there were no substantial increases in parameters related to inflammageing. Age was a more important factor in inflammation status, notably in the kidneys, while the early-life dietary switch may have influenced intestinal susceptibility to inflammation. This study affirms the therapeutic potential of RAGE modulation and corroborates evidence for the disruptive effect of dietary changes occurring too early in life. Future research should prioritize the potential influence of dAGEs on disease aetiology and development, notably any exacerbating effects they may have upon existing health conditions.

Type 2 diabetes (T2D) constitutes a major public health problem, and despite prevention efforts, this pandemic disease is 'one of the deadliest diseases in the world. In 2022, 6.7 million T2D patients died prematurely from vascular complications. Indeed, diabetes increases the risk of myocardial infarction or stroke eightfold. The identification of the molecular actors involved in the occurrence of cardiovascular complications and their prevention are therefore major axes. Our hypothesis is that factors brought into play during physiological aging appear prematurely with diabetes progression. Our study focused on the aging of the extracellular matrix (ECM), a major element in the maintenance of vascular homeostasis. We characterized the morphological and functional aspects of aorta, with a focus on the collagen and elastic fibers of diabetic mice aged from 6 months to non-diabetic mice aged 6 months and 20 months. The comparison with the two non-diabetic models (young and old) highlighted an exacerbated activity of proteases, which could explain a disturbance in the collagen accumulation and an excessive degradation of elastic fibers. Moreover, the generation of circulating elastin-derived peptides reflects premature aging of the ECM. These extracellular elements contribute to the appearance of vascular rigidity, often the origin of pathologies such as hypertension and atherosclerosis. In conclusion, we show that diabetic mice aged 6 months present the same characteristics of ECM wear as those observed in mice aged 20 months. This accelerated aortic wall remodeling could then explain the early onset of cardiovascular diseases and, therefore, the premature death of DT2 patients.

Background The approximately fifteen-year reduction in life expectancy observed in diabetic patients, compared to non-diabetic individuals, is believed to be attributed to the early onset of cardiovascular diseases. Among the molecular actors involved in the occurrence of cardiovascular complications, the remodeling of elastic fibers (EFs) in favor of degradation rather than neosynthesis is significant. Objective This study aims to modulate the elastogenesis/elastolysis balance in the arterial wall of diabetic db/db mice (a diabetic model where the leptin receptor is deficient) to limit the premature aging of their EFs and aortic stiffening. Methods Mice are treated with two antihypertensive agents: minoxidil (an ATP-sensitive potassium (KATP) channel opener) or nebivolol (a β-blocker also active on KATP channels). The degree of wear and functionality of EF are assessed after these treatments. We complement this analysis by identifying molecular actors from smooth muscle cell cultures. Results Our data show that by applying these antihypertensive agents in cultured vascular smooth muscle cells in vitro and in diabetic mice, we efficiently stimulate elastogenesis and inhibit elastolysis. Therefore, treatments restore functional EFs and limit their degradation. This brings blood pressure values of diseased mice close to normal ones (as in unaffected mice). Elastogenesis pathway stimulation and elastolysis inhibition are induced by the opening of sensitive KATP channels and the regulation of the forkhead box transcription factor (FOXO1). Conclusion Monitoring these two pathways could, therefore, be sufficient to limit the premature aging of the aorta and to reduce the occurrence of hypertension, atherosclerosis, and aneurysms in diabetic patients.

Fibroblast growth factor 23 (FGF23) is associated with cardiovascular events in patients with chronic kidney disease (CKD). Prognotic value of FGF23 in patients with cardiovascular disease (especially after acute myocardial infarction [MI]) without CKD remains unknown. To assess FGF23 levels in patients after acute myocardial infarction and explore if elevated FGF23 levels are associated with impaired survival. All patients included in the MAVASCOR study (Markers of Vascular Aging and Coronary Syndromes, NCT02857387) were included for plasmatic FGF23 assay at baseline (M0) and 12 months after MI (M12). Main endpoint was FGF23 levels at M0 and M12, obtained by in vitro quantitative measurement by immunoassay (FGF-23 Kit ELISA, ABIN1114915). High FGF23 levels were defined as FGF23≥60 pg/mL. Secondary endpoints were 1-year major adverse cardiovascular events (MACEs) defined as all-cause death, acute coronary syndrome, heart failure hospitalization or coronary revascularization. Among the 64 patients included, mean age was 56 ± 11 years, 81% were males and 4 (6.7%) presented MACEs at one year (Table 1). Less than 10% of patients had impaired renal function (defined by estimated glomerular filtration rate [eGFR] < 60 mL/min/1.73 m2). Mean FGF23 levels at M0 and M12 were 61.7 ± 91.5 and 166.6 ± 451.6 pg/mL respectively and did not differ significantly after t-paired test (p = 0.13). One fifth of patients had high FGF23 levels (≥ 60 pg/mL): 14 (22%) at M0 and 13 (20%) at M12. In univariate logistic regression, high baseline FGF23 levels were significantly associated with 1-year MACEs, compared to normal pFGF-23 (OR 13.4, 95% CI [1.27–140.93]; p = 0.031). Similar result was found after adjustement on baseline eGFR (p = 0.024). After linear regression using inverse function (for normalization of FGF23, see Figure 1A and 1B), baseline FGF23 levels were significantly associated with body mass index (p = 0.01) and NT-proBNP before and after adjustement on eGFR (p = 0.008 and 0.006 respectively; Figure 1C). No association was found between FGF23 levels and other markers of vascular aging explored in the MAVASCOR study. After acute myocardial infarction, 20% of patients had elevated FGF23 levels, independently from renal function. Early elevation of FGF23 levels after MI is associated with 1-year major cardiovascular events. Further studies are required to explore these findings.

Stiffening of the vascular network is associated with the early stages of vascular aging, leading to cardiovascular disorders (hypertension), renal failures, or neurodegenerative diseases (Alzheimer's). Unfortunately, many people remain undiagnosed because diagnostic methods are either unsuitable for a large population or unfamiliar to clinicians which favor the hypertension evaluation. In preclinical research, stiffness studies are often partially conducted. We think that the evaluation of aortic stiffness is essential as it would improve our understanding of aging diseases progression. We propose here a systematic method using decision trees in a multi-scale and multimodal approaches. Our method was evaluated by analyzing the aortic situation in old and young mice. We demonstrate that both the endothelial and smooth muscle cells exhibit pronounced functional alterations in favor of constriction. Additionally, there is significant remodeling of the extracellular matrix, leading to a drastic degradation of elastic fibers and the accumulation of collagen in the aortic wall. This series of changes contributes to the development of vascular rigidity, a preliminary stage of arterial hypertension. Our results suggest that our method should improve preclinical understanding and encourage clinicians to equip themselves with tools for assessing vascular function, as it is an essential issue for preventing numerous pathologies.

Introduction: Carbamylation is a nonenzymatic post-translational modification of proteins characterized by the binding of isocyanic acid to amino groups of proteins, which leads to the alteration of their properties. An increase in serum carbamylation-derived products, including homocitrulline (HCit), has been shown to be associated with the development of cardiovascular diseases. Methods: HCit was quantified by LC-MS/MS within extracts of aneurysmal and control human aortas. A mouse model of aortic aneurysm (ApoE−/− mice perfused with angiotensin II and fed with sodium cyanate) was used to evaluate the role of carbamylation in aneurysm development. Results: HCit quantification showed a greater heterogeneity of values in aneurysmal aortas in comparison with control ones. At the maximum diameter of dilation, HCit values were significantly higher (+94%, p < 0.05) compared with less dilated areas. No differences were observed according to aneurysm size or when comparing ruptured and unruptured aneurysms. No significant effect of carbamylation on aneurysm development was observed using the animal model. Conclusions: These results evidenced the accumulation of HCit within aneurysmal aortas but do not allow concluding about the exact participation of protein carbamylation in the development of human abdominal aortic aneurysms.